DEPC-treated BSA? - (Mar/19/2008 )
Hi all,
I have some troubles to deal with BSA.
Well, I'm doing kinetic expts on RNase A and the substrate is single strand DNA. I have to avoid the RNase/DNase contamination from the environment. Meanwhile BSA is added to stabilize the enzyme RNase A. It's okay to use the commercial RNase/DNase-free BSA soln but it's kind of expensive. I bought RNase/DNase free BSA solid but it seems there is still residual RNase/DNase.
I wonder if BSA can be DEPC-treated although I know DEPC can't be applied to compounds w/ -NH2.
Please give me some hints if you know something about this.
thanks a lot!
I have some troubles to deal with BSA.
Well, I'm doing kinetic expts on RNase A and the substrate is single strand DNA. I have to avoid the RNase/DNase contamination from the environment. Meanwhile BSA is added to stabilize the enzyme RNase A. It's okay to use the commercial RNase/DNase-free BSA soln but it's kind of expensive. I bought RNase/DNase free BSA solid but it seems there is still residual RNase/DNase.
I wonder if BSA can be DEPC-treated although I know DEPC can't be applied to compounds w/ -NH2.
Please give me some hints if you know something about this.
thanks a lot!
Are you making your BSA up with DEPC-treated water? I don't know about DEPC treating the BSA once it's already made because usually one would autoclave the DEPC treated solution in order to kill the DEPC, and you probably wouldn't want to autoclave your BSA.
Yes, i made the BSA soln w/ DEPC-treated water. But this BSA soln is not RNase-free. So I wonder if I can do anything to get rid of the RNase/DNase in the BSA.
as long as you don't use it as a standard for protein determination (and probably for size) you can use acetylated bsa (it may be a little expensive):
sigma-aldrich acetylated bsa
You might just want to use the BSA solution that NEB supplies. If you read the FAQ about it, it is clear that they have had issues over the years with nuclease contamination, first trying to solve them with acetylation, and finally switching to an (unspecified) heat treatment.